There are over three million needle biopsies performed every year in the USA, typically for the diagnosis of breast, prostate, thyroid, lung, liver, and pancreatic cancers. Because needle biopsies are less invasive than surgical biopsies, they are much more cost-effective. However, a major limitation for all needle biopsies are that the pathologist or cytologist views the tissue or cells in only two dimensions, while the important structures for determining cancer and invasiveness of the disease is more clearly seen in threes dimensions (3D) versus two dimensions. What is true in radiology is expected in pathology, 3D imaging will give a quantum increase in the information content, expected to increase sensitivity and specificity of early cancer diagnosis. For many cases such as suspected pancreatic cancer, the small needle biopsy is all the tissue that the clinical team has to test before making a lifesaving decision. In this proposal, the preferred specimen for pancreatic cancer diagnosis is a thin needle core biopsy, which the entire 1-2cm long specimen is visualized in 3D using advanced optical imaging. After needle procurement from ex vivo pancreas, all handling and preparation for highest quality 3D imaging will be prototyped and validated within a novel microfluidics device. Every tissue preparatory step of fixing, staining, clearing, and transporting to 3D imaging is conducted within the microfluidics channel, which can be automated. The goal of this project is to reduce the time for biopsy tissue preparation to less than half the standard time in a pathology lab. Furthermore, the 3D architecture of the tissue will be preserved for the first opto-mechanical measures of biopsy intactness. The diagnostic value of the needle biopsy specimens after automated preparation will be determined by experienced pathologists. The ability to visualize 3D morphology within tissue biopsies will also allow the pathologists to bette compare ex vivo diagnosis with new in vivo 3D imaging. These ex vivo technologies that rely on optically cleared tissue and traditional bright-field imaging (Z-stack imaging and optical projection tomographic microscopy) can become the pathologists' bridge for better understanding in vivo 3D imaging technologies, such as confocal, optical coherence tomography, and photo-acoustic imaging. This project is expected to lead to a transformation in pathology from 2D to 3D, and in the ability to provide less invasive, low-cost and rapid cancer diagnosis, directly affecting several millions of US citizens per year.